Multiplex characterization of human pathogens including species and antibiotic-resistance gene identification. Issue 1 (January 2016)

• Record Type: Journal Article
• Authors: Barišić, Ivan; Petzka, Josefine; Schoenthaler, Silvia; Vierlinger, Klemens; Noehammer, Christa; Wiesinger-Mayr, Herbert
• Is Part Of: Journal of medical microbiology. Volume 65:Issue 1(2016)
• Publication Date: 2016-01
• Subjects: Medical microbiology -- Periodicals; 616.9041
• DOI: 10.1099/jmm.0.000192
• Languages: English
• ISSNs: 0022-2615

Abstract

Abstract : The efficient medical treatment of infections requires detailed information about the pathogens involved and potential antibiotic-resistance mechanisms. The dramatically increasing incidence of multidrug-resistant bacteria especially highlights the importance of sophisticated diagnostic tests enabling a fast patient-customized therapy. However, the current molecular detection methods are limited to either the detection of species or only a few antibiotic-resistance genes. In this work, we present a human pathogen characterization assay using a rRNA gene microarray identifying 75 species comprising bacteria and fungi. A statistical classifier was developed to facilitate the automated species identification. Additionally, the clinically most important β-lactamases were identified simultaneously in a 100-plex reaction using padlock probes and the same microarray. The specificity and sensitivity of the combined assay was determined using clinical isolates. The detection limit was 10 5 c.f.u. ml − 1, recovering 89 % of the detectable β-lactamase-encoding genes specifically. The total assay time was less than 7 h and the modular character of the antibiotic-resistance detection allows the easy integration of further genetic targets. In summary, we present a fast, highly specific and sensitive multiplex pathogen characterization assay.